A manual of rice seed health testing - IRRI books - International Rice ...

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CHAPTER 3 Equipment J.K. Misra, T.W. Mew, and S.D. Merca Steps followed for the phytosanitary certification of rice seeds are shown in Figure 3.1. Seed testing laboratories use seed sampling equipment to draw and divide official seed samples (Table 3.1). Standard plant pathology laboratory equipment and facilities suitable for many techniques used in rice seed health testing are listed in Tables 3.1 and 3.2 and briefly explained below. The equipment used for sampling is described in the next chapter. A hand or magnifying lens (Fig. 3.2) may detect plant debris, sclerotia, smuts, malformations, and indications of infection such as dried bacterial ooze, resting mycelia on seed surfaces, and fruiting bodies. A stereobinocular microscope (Fig. 3.3) can readily detect contamination of the seed surface with teleospores of Tilletia barclayana. Additional equipment is needed for the washing test. An automatic shaker standardizes shaking, a lowspeed centrifuge concentrates suspension, a compound microscope helps identify fungal spores, and a haemocytometer determines the spore load. Stereobinocular and compound microscopes are used to detect and identify nematodes in suspension after sedimentation. 3.1 Steps toward the phytosanitary certification of rice seeds.
The blotter test, an indirect method of major importance, requires near-ultra violet (NUV) light incubation chambers equipped with timers to regulate NUV for 12 h, followed by 12 h darkness, daily. The incubation chamber temperature for the standard blotter test is 20±2 °C. Some NUV chambers are constructed inside a walk-in room that is air-conditioned to maintain the recommended temperature. The NUV light source should be a 320- 400 nm lamp, preferably Philips TL 40W/08, GE F 40 BL, or equivalent. A freezing blotter may be used to kill the germinating embryo or seedling when germination is so vigorous that seedling growth elevates the petri dish cover and the blotter becomes dry. Freezing is accomplished on the second or third day of incubation by placing the seeded plates in a freezer for 24 h and then returning them to NUV incubation. Plastic petri dishes are recommended as they allow better NUV light penetration than do glass dishes. Agar for fungi test should be incubated at 28±1 °C in NUV light/ darkness, and that for bacteria at 28 °C. This can be done in standard laboratory-top incubators. For the growing-on test, container covers for the medium (substrate) and test seeds should be transparent to admit light and to facilitate visual examination without removing the covers. Transparent plastic bags supported by sticks or ready-made hard transparent covers used in germination tests are available. Table 3.1. Laboratory equipment, instruments, and glassware used for seed health testing. a Seed sampling Stick or sleeve trier (Nobbe trier. Fig. 4.1) Conical divider (Boerner type, Fig. 4.3) Soil divider (Riffle type, Fig. 4.4) Centrifugal divider (Gamet type. Fig. 4.5) Magnifying lens (Fig. 3.3) Seed health testing Essential Autoclave Sterilizing oven Hot air Refrigerator/freezer Clean bench or laminar flow hood Centrifuge (up to 20,000 rpm) Centrifuge (table top model - 3,000-5,000 rpm) Automatic shaker or rotary shaker (adjustable speed) Distilling apparatus Incubation chambers (25-32 °C) NUV incubator (21 °C) with automatic time switch for lamps Lighted incubator (25-32 °C) Stereobinocular microscope - magnification up to 50X Compound microscope - magnification up to 1000X Mineral light lamp-UV Hand lens Haemocytometer Tally counter Mortar and pestle Electric mixer Vortex mixer Hot water bath pH meter Alcohol lamp Optional Growth chamber Hot plate Filtration apparatus Gas burner Electrostatic air cleaner Phase-contrast microscope Glassware and other laboratory supplies Petri dishes - glass, plastic Test tubes - assorted sizes Beakers - assorted sizes Syringes - assorted sizes Erlenmeyer flasks - assorted sizes Atomizer or sprayer Baermann funnel set-up (Fig. 8.1) Funnels - glass, plastic Graduated cylinders - 50, 100, 500, 1000 ml Pipettes - assorted sizes Germination trays with transparent covers Glass rods, L-shaped Microscope slides/cover glasses Filter paper Blotters, round, 8.5 cm diameter Wire loop (transfer loop) Inoculating needle Plastic bags - different sizes Mineral oil or paraffin Marking pens for glass Indelible pencils Paper towels Working box containing dissecting needles, forceps, scissors, scalpels, transfer needles, etc. a Sources: IRRl Seed Health Unit and Langerak et al (1988). 14 Rice seed health testing manual
Page 2 and 3: A Manual of Rice Seed Health Testin
Page 4 and 5: Contents Foreword v Preface vi Part
Page 6 and 7: Foreword Intensive collaboration ch
Page 8: Part 1 Introduction CHAPTER 1 Rice
Page 11 and 12: tional levels (Kahn 1988). Quaranti
Page 13 and 14: Morphology of the mature plant Matu
Page 15 and 16: two-celled, besifixed, deeply sagit
Page 18 and 19: Part 2 Procedures for seed health e
Page 22: 3.2 Dry seed inspection using a mag
Page 25 and 26: Example. In a two-stage random samp
Page 27 and 28: solid pointed end. The tube and sle
Page 30: CHAPTER 5 Dry seed inspection J.K.
Page 33 and 34: Detection Blotter and agar plate me
Page 35 and 36: Results: Examine plates for charac-
Page 37 and 38: Extraction and isolation It is esse
Page 39 and 40: GRINDING METHOD (FOR SMALL-SCALE SE
Page 41 and 42: flagellum. Cells occur singly, in p
Page 43 and 44: 7.6 Oxidase test reaction. L = colo
Page 45 and 46: 7.9 Production of 2-ketogluconate.
Page 47 and 48: 1. Grow pathogen-free plants under
Page 49 and 50: 7.12 a. P. glumae type A colony on
Page 51 and 52: A1. Macerate 100 seeds in 10 ml of
Page 53 and 54: Results: 7.Streak the bacteria onto
Page 55 and 56: Sieve method (Fig. 8.1) Procedure:
Page 58 and 59: CHAPTER 10 Field inspection J.K. Mi
Page 60 and 61: Table 10.2. continued Disease Fungi
Page 62 and 63: Table 10.2. continued Disease Growt
Page 64: additional precaution, the multipli
Page 67 and 68: terracoat (5-ethoxy-3- trichloromet
Page 70:
Part 3 Pests and pathogens CHAPTER
Page 73 and 74:
elevant tests for determining the i
Page 75 and 76:
Table 12.1 continued Family and spe
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Cryptolestes pusillus Schonherr The
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Rice weeviI Scientific name: Sitoph
Page 82 and 83:
CHAPTER 14 Fungal pathogens J.K. Mi
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THE DISEASE—BROWN SPOT Sir John W
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Symptoms Glumes are discolored, and
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y low relative humidity. Released a
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THE DISEASE—STEM ROT Stem rot was
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THE DISEASE—SHEATH BLIGHT Sheath
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Sarocladium oryzae is seedborne. Th
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Although not currently considered a
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Pseudomonas fuscovaginae Pathogen:
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See Chapter 7 and Figure 15.4a for
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Infected seeds and contaminated wat
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For further details, see the Common
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nonfluorescent) and Erwinia herbico
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Franklin M T, Siddiqi M R (1972) Ap
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Tai F L, Siang W N (1948) 'I-chu-hs
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106 Rice seed health testing manual
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108 Rice seed health testing manual
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After autoclaving the medium, add t
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smoniloid = having swellings at reg
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